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- [Protocol]Construction of a multicopy genomic DNA library and its application for suppression analysis
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Hongbaek Cho
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J. Microbiol. 2019;57(12):1041-1047. Published online November 22, 2019
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DOI: https://doi.org/10.1007/s12275-019-9417-8
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Abstract
- Suppression analysis is used for the identification of new genes
and genetic interactions when there is a notable phenotype
available for genetic selection or screening. A random genomic
DNA library constructed on a multi-copy plasmid is a
useful tool for suppression analysis when one expects that
an overdose of a few genes will suppress the phenotype. These
libraries have been successfully used to determine the function
of a gene by revealing genes whose functions are related
to the gene of interest. They have also been used to identify
the targets of chemical or biological agents by increasing the
number of unaffected target gene products in a cell. In this
article, I will discuss important considerations for constructing
multicopy genomic DNA libraries. The protocol provided
in this paper should be a useful guide for constructing
genomic DNA libraries in many bacterial species for which
multi-copy plasmids are available.
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